129s6 svev stat1tm1rds

C57BL/6, OT-I transgenic (C57BL/6-Tg(TcraTcrb)1100Mjb/J), RIP-mOVA (C57BL/6-Tg (Ins2-TFRC/OVA)296Wehi/WehiJ) mice, Rorc^−/− (B6.129P2(Cg)-Rorctm2Litt/J) and Tbx21^−/− (B6.129S6-Tbx21tm1Glm/J) mice were purchased from The Jackson Laboratory (Bar Harbor, ME). STAT1-deficient mice (129S6/SvEv-Stat1tm1Rds) were purchased from Taconic Farms (Cranbury, NJ). The Institutional Animal Care and Use Committee of Thomas Jefferson University approved all experimental procedures.

The mouse line
B6.129S7-Rag1^tm1Mom/J
(referred to as
Rag1^−/−) and
control animals of the C57BL/6J strain were obtained from The Jackson
Laboratory (Bar Harbor, ME). The mouse line
129S6/SvEv-Stat1^tm1Rds(referred to as
Stat1^−/−)and control animals of the 129S6/SvEvTac strain (referred as 129S6) were
obtained from Taconic (Rensselaer, NY). For all animal experiments, 6-to
8-week-old female mice were randomly assigned to experimental groups and
housed under Specific Pathogen Free (SPF) conditions. All animal studies
were reviewed and approved by the Institutional Animal Care and Use
Committee (IACUC) of the Icahn School of Medicine at Mount Sinai, in
accordance with the institutional and national guidelines and regulations
and performed in Association for the Assessment and Accreditation of
Laboratory Animal Care (AAALAC)-certified facilities.

Six week old 129Sv WT mice and Stat1 knockout (KO) mice (129S6/SvEv-Stat1^tm1Rds) were purchased from Taconic (Germantown, NY). All animals were maintained under specific pathogen-free conditions and used in strict accordance with the guidelines and protocols approved by The Johns Hopkins University School of Medicine Animal Care and Use Committee. Experiments were repeated 3 times; experiment 1 used 20 WT and 25 Stat1 KO mice; experiment 2 used 30 WT and 30 Stat1 KO mice, and experiment 3 used 19 WT and 19 Stat1 KO mice.
Low passage (p4) A. phagocytophilum Webster strain was used. On the day of inoculation, 10⁴ heavily infected (>90%) HL-60 cells were centrifuged (2,000 × g, 10 min) to concentrate the cells. Cell-free A. phagocytophilum was isolated by syringe lysis, cellular debris removed by centrifugation at 3,000 × g for 10 min, and the host cell-free A. phagocytophilum were then harvested from the supernatant by centrifugation at 12,000 × g for 30 min. Bacteria were resuspended in PBS for experimental infections and injected intraperitoneally into mice. PBS alone was used for mock infections. Five mice of each strain were assigned to A. phagocytophilum-infected and mock-infected cohorts for evaluation at time intervals as follows: 4 h, 2, 4, 7, 10, and 14 days post infection. All experiments were repeated twice to confirm reproducibility.

C57BL/6 (B6), B6.129S2-Irf1^tm1Mak/J, B6.129S2-Batf^tm1.1mm/J and B6.SJL-PtprcaPepcb/BoyJ (CD45.1) were purchased from the Jackson Laboratories. B6.129S6-Rag2tm1Fwa N12, 129S6/SvEv-Stat1^tm1Rds and 129S6/SvEvTac controls were purchased from Taconic. Animals were maintained in a conventional, pathogen-free facility at the Harvard Institutes of Medicine (Boston, MA) and all experiments were carried out in accordance with guidelines prescribed by the Institutional Animal Care and Use Committee (IACUC) at Harvard Medical School.